The hepatotoxicity of carbon tetrachloride is usually thought to be due to the enzymatic formation of the trichloromethyl radical. A variety of indirect, but not conclusive, evidence for the formation of .CCl3 exists: hydrogen abstraction by .CCl3 to form CHCl3 and dimerization of .CCl3 to form C2Cl6. Hydrogen abstraction of a methylene hydrogen from polyunsaturated fatty acids by the trichloromethyl radical would be followed by oxygen addition and would result in lipid peroxidation. Carbon tetrachloride-induced lipid peroxidation has been extensively studied both in vitro and in vivo. Attempts to use electron spin resonance (ESR) spectroscopy to demonstrate directly the presence of the trichloromethyl radical in hepatic microsomal incubations or liver slices have been unsuccessful. Recently a free radical has been detected in microsomal incubations containing NADPH and CCl4 or CBrCl3 using the spin-trap phenyl-t-butyl nitrone (PBN). This free radical adduct was identified as the .CCl3 adduct of PBN. Our studies have shown, however, that a lipid dienyl radical, similar to that formed by the action of soybean lipoxygenase on linoleic acid, is the species being trapped.